1. Field of the Invention
The present invention is directed to assay apparatus, devices and methods that permit longitudinal capillary flow of liquid between two pieces of bibulous material and that, prior to actuation, are in a non-capillary flow relationship to each other. In particular, the device utilizes a distortable member which when distorted, actuates the device and a capillary flow relationship is initiated between the two pieces of bibulous material. The methods, apparatus, and devices of the present invention have utility in conducting assays which use capillarity to transport solutions and/or samples including assays such as enzyme immunoassays, fluorescent immunoassays, radioimmunoassays, etc.
2. Related Art
Assay devices have been described in which liquid is transported by capillarity through or transversely along a bibulous support thereby transporting reagents and samples to sites on the support and/or washing the support. Such devices have been described by Deutsch, U.S. Pat. No. 4,094,647; Zuk et al, U.S. Pat. No. 4,435,504; Weng et al, U.S. Pat. No. 4,740,468; Friesen et al, German Offenlengungschrift 3,445,816 etc.
In many of these methods, reagents can be added at more than one position on the bibulous support. In such situations, it is often desirable to cause the added reagent to migrate in only one direction along the bibulous support, to delay flow in a given direction, or to create a direction of flow that was impeded prior to the addition of the reagent. For example, it is often desirable for a solution moving along a bibulous support to automatically stop moving after a certain volume has been taken up so as to avoid the necessity to monitor the flow of solution. Subsequent to taking up a certain volume, a second solution is allowed to flow along the support. In the past, the first flow could be stopped by limiting the length of the first bibulous support and the second flow initiated by contacting one end of the first bibulous support with a solution and the other end with a second bibulous support which is dry.
Likewise, in liquid containing assay devices, a sample is added to one of the ports and liquid within the device washes the sample away from a detection zone. See, for instance, Khanna et al, U.S. patent application Ser. No. 35,562 filed Apr.7, 1987 entitled "Immunoassay Devices". To maximize the washing efficiency, it is desirable that upon contacting the support with the sample, the sample flows in only the direction away from the source of the wash liquid.
In both of the above cases, it is desirable to create a capillary flow relationship between pieces of bibulous material which heretofore have been in a non-capillary flow relationship. Moreover, it would be particularly desirable to create such a capillary flow relationship automatically without mechanical means external to the device while requiring minimal operator involvement to operate.
U.S. Pat. No. 3,482,943 discloses expandable sponges useful in transporting solution to a set position on a gel suitable for conducting immunodiffusion tests. In this reference, the solution transported by the sponge is allowed to diffuse into the gel which, in order to conduct the immunodiffusion, is by necessity a wet gel. Accordingly, no capillary flow relationship between the expandable sponge and the gel is established by this device.
U.S. Pat. No. 4,246,339 discloses a device having an upper portion and a lower portion. The upper portion has a plurality of wells wherein the bottom of each well is fitted with a membrane layer capable of transporting liquid. The bottom portion of the device contains absorbent material. Between the top and bottom portions is a compressible spacer. This device allows a liquid sample to be added to the wells which may optionally be impregnated with an antibody. After a set incubation period, pressure is placed on the top portion which because of the compressible spacer, allows the bottom of each of the wells to contact the absorbent material whereupon the liquid in the wells is transferred to the absorbent material. After liquid transfer, the pressure is removed and contact between the bottom of the wells and the absorbent material is broken. Thereupon, additional liquid may be added to the wells. In this device, care must be taken to insure that the bottom of all of the wells come into contact with the absorbent material for a sufficient period of time to remove the liquid. In particular, if one or more of the wells does not contact the absorbent material, then the liquid in that well will not be removed. Moreover, if one or more of the wells does not contact the absorbent material for a sufficient period of time, then not all of the liquid in that well will be removed. In any case, a high level of operator care is required to ensure the proper operation of this device.
U.S. Ser. No. 35,562 filed Apr. 7, 1987 entitled "Immunoassay Devices" discloses a device for conducting an assay method. The device comprises a housing having in one portion thereof a breakable capsule and in another portion a piece of bibulous material attached to an absorbent pad.
European Patent Application Publication No. 0 146 691 discloses an air bleed passage in a liquid sampling needle which is formed between a housing and a sleeve and into which is added a solid compacted material which swells on contact with liquid. As the liquid sample is drawn into the needle, air can escape via the air bleed until the air bleed is contacted with liquid whereupon it expands and forms a liquid impermeable membrane.
U.S. Pat. No. 4,700,741 discloses a urine collecting device which contains an expandable sponge in a compartment which permits the collection of a predetermined quantity of urine by limiting expansion of the sponge within the compartment.
Canadian Patent No. 1,185,882 discloses porous, hydrophilic, non-gel-forming swellable polymers or self-drawing fluid reservoirs with a very high and uniform absorption and release of fluid in a chromatographic quick-test device.
U.S. Pat. No. 4,826,759 describes apparatuses and methods, which can be used in the field (i.e., outside the laboratory environment) to determine qualitatively and at least semiquantitatively the presence or absence of minute quantities of ligand. The apparatus can be in the form of a strip comprising a support means provided with a groove intermediate its ends forming a crease line upon which the strip can be folded upon itself with bibulous elements and spaced from the crease line and arranged so that when the strip is folded upon itself the bibulous elements become aligned with each other and come into liquid contact.
U.S. Pat. No. 4,803,170 discusses an immunoassay device including one or more reaction chambers, each adapted to receive and retain a volume of test fluid communication with non-overlapping first, second, and third reagent-bearing surfaces. To the first surface is reversibly bound an analyte conjugate: Analyte component conjugated to one or more components, termed ligand/marker, that serve ligand and marker functions as described herein. Analyte binding partner is immobilized on the second surface, and ligand/marker binding partner is immobilized on the third surface. The reaction chamber is preferably configured to receive and direct the test fluid sequentially past the first, second, and third reagent surfaces. In use, analyte conjugate solubilized from the first surface competes with any analyte in the test fluid for analyte binding partner sites on the second surface. Excess analyte conjugate becomes sequestered on the third surface, where the marker activity is read to indicate analyte presence and concentration in the test fluid. A test kit includes the immunoassay device in combination with comparative test results.
U.S. Pat. No. 3,888,629 describes a reaction cell for the performance of radioimmunoassay determinations and like saturation analysis reactions that has supported within it a matrix pad of absorbent material capable of retaining the necessary reagents for the reaction and serving as a site in which the reaction totally occurs. A separable lower chamber is fitted to the lower end of the cell and contains absorbent material to abut the matrix pad and promote filtration through the pad after the reaction has taken place. An upper reservoir chamber fits to the upper end of the cell to contain liquid for passing through the matrix pad. The matrix pad will commonly contain prior to the reaction a predetermined amount of an antigen or antibody in freeze-dried condition and possibly radioactivity labelled.
Accordingly, there is a need for a device for creating a capillary flow relationship between pieces of bibulous material which prior to actuation, are in a non-capillary flow relationship to each other. Preferably, such a device should require minimal operator involvement to operate.